摘要
目的分析环境噪声与一氧化碳(carbon monoxide,CO)在独立及联合暴露状态下对大鼠精子发生过程的损害效应,探究可能的损伤机制及紫檀芪(pterostilbene,PTE)对该损伤的保护作用。方法采用随机数字表法将60只雄性SD大鼠[6~8周龄,体质量(200±10)g]分为6组(n=10):正常对照组(常规饲养)、假暴露对照组(单纯束缚应激)、噪声暴露组(85 dB声压级)、CO暴露组(460 mg/m^(3))、联合暴露组(同步施加460 mg/m^(3)CO与85 dB噪声)及PTE干预组[在联合暴露基础上予PTE(80 mg/kg)预处理]。实验采用动态口鼻暴露系统,整体置于多因素环境暴露模拟舱内,每天暴露2 h,持续60 d。染毒结束后分析精子数量和存活率,用HE染色观察大鼠睾丸病理组织学损伤情况,采用qRT-PCR检测不同阶段生殖细胞特异性mRNA相对水平,并检测血清性激素水平、睾丸及精子三磷酸腺苷(adenosine triphosphate,ATP)水平,通过透射电镜观察精母细胞超微结构损伤,睾丸组织进行转录组测序后行生物信息学分析。结果与假暴露对照组相比,联合暴露组精子存活率和数量显著降低(P<0.05),且病理结果提示联合暴露组大鼠睾丸组织出现未成熟生殖细胞的发育阻滞;qRT-PCR结果提示联合暴露组大鼠精原细胞、精母细胞以及圆形和长形精子细胞数量均显著降低(P<0.01);联合暴露组生殖相关激素(如促性腺激素释放激素、卵泡刺激素、黄体生成素和睾酮)以及睾丸和精子ATP水平均显著降低(P<0.01),且透射电镜观察显示精母细胞线粒体破裂和嵴破坏。与联合暴露组相比,PTE干预组上述损伤效应有明显好转,并趋于正常水平。转录组测序结果提示,生殖发育以及ATP依赖的相关生物过程可能参与噪声和CO暴露对大鼠睾丸的损伤效应,其中关键基因包括Nppa、Adm、Gnrh1、Ptafr、Atp13a5、Atp8b1、LOC102555469等。结论噪声和CO可导致大鼠精子发生过程损伤,其机制可能与能量代谢和激素有关,且PTE可显著改善该生殖损伤。
Objective To analyze the detrimental effects of exposure to environmental noise alone and combined with carbon monoxide(CO)on spermatogenesis in male rats,investigate the underlying mechanisms involved in such damage,and evaluate the protective role of pterostilbene(PTE)against these adverse effects.Methods Sixty male SD rats(6~8 weeks old,weighing 200±10 g)were randomly divided normal control group(standard housing),sham-exposure control group(restraint stress only),noise exposure(85 dB),CO exposure(460 mg/m^(3)),combined exposure(simultaneous exposure),and PTE intervention(80 mg/kg pretreatment),with 10 animals in each group.The rats were exposed daily for 2 h via a nose-only inhalation exposure system within a multifactorial environmental simulation chamber for 60 consecutive days.Sperm count and viability were measured after exposure.Histopathological changes of testicular tissues were observed with HE staining.qRT-PCR was used to measure stage-specific mRNA levels in germ cells.Serum sex hormone levels and adenosine triphosphate(ATP)concentrations in testes and sperm were detected.Transmission electron microscopy(TEM)was applied to observe the ultrastructural damage in the spermatocytes.Additionally,transcriptome sequencing was performed on testicular tissue,followed by bioinformatics analysis.Results Compared with the negative control group,the combined exposure group exhibited significant reductions in sperm viability and count(P<0.05),and developmental arrest of immature germ cells in the testicular tissue,with obviously less spermatogonia,spermatocytes,and round/elongated spermatids(P<0.01).Additionally,significantly reduced levels of reproductive-related hormones,such as gonadotropin-releasing hormone,follicle-stimulating hormone,luteinizing hormone and testosterone,and ATP levels in testes and sperm were observed in the mice after combined exposure(P<0.01),accompanied by mitochondrial rupture and cristae disruption in spermatocytes.Conversely,the PTE intervention group showed marked alleviation of these impairments,with parameters recovering almost to normal levels.Transcriptome sequencing identified biological processes related to reproductive development and ATP-dependent pathways as potential contributors to testicular injury induced by noise and CO exposure,with key genes including Nppa,Adm,Gnrh1,Ptafr,Atp13a5,Atp8b1,and LOC102555469.Conclusion Noise and CO exposure induce spermatogenic damage in rats,which may be related with energy metabolism and hormonal regulation,
作者
郭橹橹
张中豪
刘畅
师超峰
王建康
孙磊
刘晋祎
李迎庆
GUO Luu;ZHANG Zhonghao;LIU Chang;SHI Chaofeng;WANG Jiankang;SUN Lei;LIU Jinyi;LI Yingqing(Department of Military Toxicology,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,China)
出处
《陆军军医大学学报》
北大核心
2025年第15期1729-1740,共12页
Journal of Army Medical University
基金
国家自然科学基金面上项目(82173555)
国家自然科学基金重点项目(82130097)。
关键词
噪声
一氧化碳
联合暴露
雄性生殖毒性
精子发生
三磷酸腺苷
紫檀芪
noise
carbon monoxide
combined exposure
male reproductive toxicity
spermatogenesis
adenosine triphosphate
pterostilbene
